Closed-state inactivation and pore-blocker modulation mechanisms of human CaV2.2

Abstract

N-type voltage-gated calcium (Ca_V) channels mediate Ca²⁺ influx at presynaptic terminals in response to action potentials and play vital roles in synaptogenesis, release of neurotransmitters, and nociceptive transmission. Here, we elucidate a cryo-electron microscopy (cryo-EM) structure of the human Ca_V2.2 complex in apo, ziconotide-bound, and two Ca_V2.2-specific pore blockers-bound states. The second voltage-sensing domain (VSD) is captured in a resting-state conformation, trapped by a phosphatidylinositol 4,5-bisphosphate (PIP₂) molecule, which is distinct from the other three VSDs of Ca_V2.2, as well as activated VSDs observed in previous structures of Ca_V channels. This structure reveals the molecular basis for the unique inactivation process of Ca_V2.2 channels, in which the intracellular gate formed by S6 helices is closed and a W-helix from the domain II-III linker stabilizes closed-state inactivation. The structures of this inactivated, drug-bound complex lay a solid foundation for developing new state-dependent blockers for treatment of chronic pain.